Open Access Review Article Series of Botany and Environmental Science Vol 3 Iss 1 Phytochemicals in Macrotyloma uniflorum – A Review Mohanraj R Houston Community College, Houston, TX, USA *Correspondence: Remya Mohanraj, Houston Community College, Houston, TX, USA Received on 18 November 2020; Accepted on 21 January 2021; Published on 26 March 2021 Copyright © 2021 Mohanraj R. This is an open access article and is distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Abstract Macrotyloma uniflorum (horse gram), is an underutilized legume that has high nutritional value. It is highly drought resistant and the seeds possess medicinal properties by virtue of the wide array of phytochemicals harbored in them. This article attempts to present a comprehensive review of the phytochemicals present in the seeds of M. uniflorum. Information presented in this chapter has been compiled from various published sources. Keywords: horse gram, Macrotyloma uniflorum, phytochemicals, review Introduction Horse gram (Macrotyloma uniflorum (Lam.) Verdcourt (Syn., Dolichos uniflorus Lam., Dolichos biflorus auct. non L.)) [1] belonging to Fabaceae, is protein-rich and is amenable for cultivation in dry conditions and marginal soil fertility [2]. Flowers together in the leaf axils without a common peduncle [3]. Seeds of horse gram are ovoid in shape and colored pale fawn, light red, brown, or black [4]; reddish brown or grey [5]; pale brown, medium brown and blackish brown [6] (Figure 1). In ayurvedic medicine, whole seeds of horse gram, is advocated in the treatment of renal stones, piles, oedema, etc. [7]. The seeds have been reported to be a significant source of iron, molybdenum [7, 8] and protein [9]. Traditional medicinal use of horse gram seeds is emphasized by its mention in Charak Samhita (renowned text on Indian medicinal system) as a remedy for piles, hiccup, abdominal lump, bronchial asthma, for causing and regulating perspiration [10]. Kirtikar et al. in their book Indian medicinal plants, note Sanskrit writers’ recommendation of horse gram’s use as a demulcent in calculous affections cough, etc. [3]. The seeds find use in herbal medicine not only as tonic, astringent, diuretic but also as a remedy for asthma, bronchitis, urinary discharges, hiccoughs, ozoena, heart trouble and brain diseases [11]. M. uniflorum’s utilization in Indian traditional medicine in treatment of urinary stones was reported by Ravishankar et al. [12]. A soup of the seeds could be heat generating and help dilute renal stones [13]. The figure shows two traditional South Indian dishes made using these seeds (Figure 2a and 2b). It’s common knowledge in certain parts of south India to consume Kollu Rasam especially when affected by common cold (Figure 2b) (Figure 2). Citation: Mohanraj R. Phytochemicals in Macrotyloma uniflorum – a review. Series Bot Environ Sci. 2021;3(1):1-9. Series of Botany and Environmental Science 2021 | Vol 3 | Iss 1 Figure 1: Seeds of M. uniflorum. Figure 2: (a) Kollu chutney (b) Kollu rasam- South Indian dishes made using horse gram. Various studies have corroborated the anti-hypercholesterolemic effect [14], anti-microbial activity [15–17] anti- helmintic activity [18, 19] anti-inflammatory [20], anti-diabetic [21], anti-oxidant [22, 23] and anti-urolithiatic activity [13, 24–26] of M. uniflorum seeds. Auxilia et al. reported that dolichins from the horse gram plant possess anti-HIV properties [27]. From the food composition table published by The National Institute of Nutrition (ICMR) by sampling whole horse gram seeds from different geographical regions of India, the protein content of horse gram seeds is comparable to other legumes and almost twice the protein content as of most cereal grains [28]. Although rich in proteins, intake of these seeds is restricted to certain groups because of its less acceptable taste and flavor and has therefore been an underutilized food legume [29]. Keeping the above background in mind, the present chapter is aimed at presenting a comprehensive review of various published reports on phytochemicals in M. uniflorum seeds. The review intends to provide the necessary background information for scientists to carry out studies on this valuable and yet underutilized legume. Electronic literature search was conducted to look for various recent studies that had analyzed the phytochemicals from M. uniflorum. Methods of Phytochemical Analysis Phytochemicals help defend plants against biotic and abiotic stresses [30, 31]. They have varying effects as antioxidants, enzyme action modulators, immune stimulators, hormone metabolism modulators, anti-bacterial and antiviral agents [32, 33]. Extensive investigations on the use of secondary plant metabolites as a source of pharmaceutical compounds are being carried out [34]. Isolation and purification of therapeutic biomolecules from medicinal plants opens avenues for the discovery of new drugs. M. uniflorum contains several phytochemicals which could be attributed to its beneficial effects on human health. Analysis of phytochemicals could be done qualitatively or quantitatively. Preliminary qualitative analyses include various biochemical tests performed to test the presence of phytochemicals. For instance, some of the available tests include Mayer’s, Dragendorff’s reagent for alkaloids [35, 36]; test using hydrochloric acid for flavonoids [37]; frothing test for saponins [38]; Liebermann–Burchard test, Salkowski’s test for phytosterols [39, 40]; Lowry assay for proteins 2 Series of Botany and Environmental Science 2021 | Vol 3 | Iss 1 [41]; Millon’s test with acetic acid for tannins and phenolic compounds [42]; Barfoed’s test, Fehling’s test, Benedict’s test for carbohydrates [43–45]; shaking with distilled water and observing the layer of foam formed for saponins [46]; Salkowski test for terpenoids [47]; using Folin-Ciocalteu reagent for phenols [48]; alkaline reagent test for flavonoids [49]. One or more of the available methods could be chosen depending on the conditions. De Silva et al. in their review describe gas chromatography (GC), gas chromatography coupled with mass spectrometry (GC-MS), liquid chromatography (LC), high performance liquid chromatography (HPLC), high performance thin layer chromatography (HPTLC) as qualitative and quantitative techniques for screening phytochemicals from plant materials [50]. Chromatography includes a variety of separation techniques that are based on partitioning different components of a sample between a mobile phase and a stationary phase. The mobile phase is either a gas or liquid [51]. During chromatography, the components of a mixture are separated according to their difference in affinities for a stationary phase and different chromatographic techniques use different stationary phase and operational conditions [52]. Physicochemical characteristics and instrumentation availability guide the choice of chromatographic system and qualitative and quantitative information is obtained from the retention time and peak area data from the chromatogram [51]. Structure determination of the isolated compounds could be carried out using a variety of techniques including UV- visible, infrared (IR), nuclear magnetic resonance (NMR), and mass spectroscopy. Structural determination relies on a spectrum that indicates the amount of radiation absorbed after passing electromagnetic radiation through an organic molecule [53, 54]. Phytochemicals in Macrotyloma uniflorum This section discusses the various phytochemicals that have been reported mainly from the seeds of M. uniflorum. Since other reviews on the subject are available, the present review is focused on including more recent studies. Aqueous extracts of M. uniflorum were qualitatively analyzed for various phytoconstituents by Patel et al. and they reported carbohydrates, alkaloids, flavonoids, saponins, phytosterols and phenolic compounds from the extracts [55]. Studies carried out by Ojha et al. showed that germination and fermentation of M. uniflorum seeds significantly reduces anti-nutritional factors and increases bioactive components. In addition, it was found that antioxidant content decreased during soaking and increased during roasting, germination and fermentation [56]. Manikandan et al. through phytochemical screening observed alkaloids, flavonoids, phenols, tannins, saponins, glycosides, steroids, terpenoids, proteins and carbohydrates from various solvent extracts of M. uniflorum seeds. Among the various solvents, ethanolic extract was reported to be very effective followed by methanol, aqueous, chloroform and hexane extracts [57]. Venugopal et al. reported an RF value of 0.9 in TLC plates from petroleum ether, acetone, ethanol, chloroform, ethyl acetate, methanol and water extracts of M. uniflorum. The authors also reported the presence of terpenoids through UV-visible spectroscopy and qualitative analysis of acetone, chloroform, methanol and aqueous extracts [58]. Sharma et al. reported compounds like syringic acid hexoside, [6]-gingerol, tyramine, rutaecarpine, asperuloside, abscisic acid, proline betaxanthin and prostaglandin E1 from methanolic extracts of M. uniflorum through liquid chromatography mass spectrometry and proximate analysis [59]. Jaya et al. through qualitative phytochemical analysis of acetone, chloroform, ethanol and aqueous extracts of M. uniflorum seeds reported the presence of carbohydrate, protein, alkaloids, flavonoids,